Trehalose metabolism genes in Caenorhabditis elegans and filarial nematodes

被引:100
|
作者
Pellerone, FI
Archer, SK
Behm, CA
Grant, WN
Lacey, MJ
Somerville, AC
机构
[1] Australian Natl Univ, Sch Biochem & Mol Biol, Fac Sci, Canberra, ACT 0200, Australia
[2] CSIRO Entomol, Canberra, ACT 2601, Australia
[3] AgResearch, Wallaceville Anim Res Ctr, Upper Hutt, New Zealand
关键词
trehalose; trehalose-6-phosphate synthase; trehalase; RNA interference;
D O I
10.1016/S0020-7519(03)00173-5
中图分类号
R38 [医学寄生虫学]; Q [生物科学];
学科分类号
07 ; 0710 ; 09 ; 100103 ;
摘要
The sugar trehalose is claimed to be important in the physiology of nematodes where it may function in sugar transport, energy storage and protection against environmental stresses. In this study we investigated the role of trehalose metabolism in nematodes, using Caenorhabditis elegans as a model, and also identified complementary DNA clones putatively encoding genes involved in trehalose pathways in filarial nematodes. In C. elegans two putative trehalose-6-phosphate synthase (tps) genes encode the enzymes that catalyse trehalose synthesis and five putative trehalase (tre) genes encode enzymes catalysing hydrolysis of the sugar. We showed by RT-PCR or Northern analysis that each of these genes is expressed as mRNA at all stages of the C. elegans life cycle. Database searches and sequencing of expressed sequence tag clones revealed that at least one tps gene and two tre genes are expressed in the filarial nematode Brugia malayi, while one tps gene and at least one tre gene were identified for Onchocerca volvulus. We used the feeding method of RNA interference in C. elegans to knock down temporarily the expression of each of the tps and tre genes. Semiquantitative RT-PCR analysis confirmed that expression of each gene was silenced by RNA interference. We did not observe an obvious phenotype for any of the genes silenced individually but gas-chromatographic analysis showed >90% decline in trehalose levels when both tps genes were targeted simultaneously. This decline in trehalose content did not affect viability or development of the nematodes. (C) 2003 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.
引用
收藏
页码:1195 / 1206
页数:12
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